Laboratory dignosis Flashcards
Tumor markers: classification
o Enzymes o Tissue receptors o Oncogenes o Hormones o Antigens (largest class)
Tumor markers: enzymes
In certain tissues are found in blood plasma at higher levels when the cancer involves that tissue
(PAP- prostatic acid phosphatase)
Tumor markers: Tissue receptors
Proteins associated with the cell membrane.
Bind to hormones and growth factors, and therefore affect the rate of tumor growth.
May be measured in the blood, extracellular fluid, and tissue samples after biopsy
(E and P receptors)
Tumor markers: Oncogenes
Some tumor markers are the product of oncogenes, which are genes that are active in fetal development and trigger the growth of tumors when they are activated in mature cells
(BARC1/2)
Tumor markers: Hormones
- hormones that are normally secreted by the tissue in which the malignancy arises
- hormones produced by tissues that do not normally produce it
(ACTH ectopic production)
Tumor markers: Antigens
Largest class of tumor marker ➢ Oncofetal antigens (AFP) (protein that are present only in fetal development and in adult life in cancer)
➢ Carbohydrate antigens (Ca 15-3)
➢ Blood group antigens (Ca 19-9)
Tumor markers:
Factors that influence the quality of results
- High specifity, i.e. not detectable in benign diseases and healthy subjects (has no false positives);
- High sensitivity, i.e. detectable very early when only a few cancer cells are present (has no false negatives);
- Organ specificity (PSA);
- Correlation with the tumor stage or tumor mass;
- Responsible to treatment;
- Correlation with prognosis;
- Reliable prediction value.
Tumor markers: criteria (7)
- High specifity, i.e. not detectable in benign diseases and healthy subjects (has no false positives);
- High sensitivity, i.e. detectable very early when only a few cancer cells are present (has no false negatives);
- Organ specificity (PSA);
- Correlation with the tumor stage or tumor mass;
- Responsible to treatment;
- Correlation with prognosis;
- Reliable prediction value.
Tumor markers: Clinical use
To detect the tumor localization, size, course of development..
GI tumor markers: (3)
Liver (3)
- AFP
- AFP-L3%
Pancreas (4)
- Ca 19-9,
- CEA,
- Ca494,
- Ca242
Colorectal (3)
- CEA,
- CTC,
- Ca 19-9
Stomach (2)
- CEA,
- Ca 72-4
GI tumor markers, clinical significance. Interpretation of results.
LIVER (3)
AFP: ▪ Useful in diagnosing cancer and in monitoring its treatment, determine prognosis or detecting tumor recurrence. ▪ Detect liver cancer at early stage. ▪ Hepatocellular carcinoma. ▪ Germ cells tumors. ▪ Chronic hepatitis.
AFP-L3:
▪ New generation TM.
▪ Specific for liver cancer (hepatocellular carcinoma)
▪ Used for screening
▪ Found to be useful indicator of distant metastasis and poor prognosis for HCC.
AFP-L3%:
▪ Very specific for small HCC.
▪ AFP-L3%>35%- 100% specificity for HCC.
Gastrointestinal tumor markers, clinical significance. Interpretation of results.
PANCREAS (2)
Ca 19-9:
▪ Not used for screening
▪ Pancreatic cancer and hepatobilliary carcinoma.
▪ Is useful in monitoring recurrence or early relapse.
Carcinoembryionic antigen (CEA):
▪ Non-specific.
▪ Not used for screening.
▪ Used for monitoring of the treatment response and reoccurrence.
▪ Colorectal cancer
▪ Pancreatic carcinoma
▪ Gastric carcinoma
▪ Lung carcinoma.
Gastrointestinal tumor markers, clinical significance. Interpretation of results.
COLORECTAL (1)
Carcinoembryionic antigen (CEA): ▪ Non-specific. ▪ Not used for screening. ▪ Used for monitoring of the treatment response and reoccurrence. ▪ Colorectal cancer ▪ Pancreatic carcinoma ▪ Gastric carcinoma ▪ Lung carcinoma.
Gastrointestinal tumor markers, clinical significance. Interpretation of results.
STOMACH (1)
Carcinoembryionic antigen (CEA): ▪ Non-specific. ▪ Not used for screening. ▪ Used for monitoring of the treatment response and reoccurrence. ▪ Colorectal cancer ▪ Pancreatic carcinoma ▪ Gastric carcinoma ▪ Lung carcinoma.
Genitourinary tract tumor markers
PROSTATE (7)
- PSA
- fPSA
- CTC
- PAP
- PSMA
- PCA3
- EPCA-2
Genitourinary tract tumor markers
TESTICULAR GERM CELL (5)
- hCG,
- fßHCG
- AFP
- LDH
- PAP
Genitourinary tract tumor markers
URINARY BLADDER (7)
- NMP22
- BTA
- CEA
- Ca 125
- Ca 19-9
- TPA
- p53
Genitourinary tract tumor markers, clinical significance. Interpretation of results.
PROSTATE (6)
➢ PSA-
▪ Used for screening, follow up and monitor of treatment.
▪ Race depended.
▪ To avoid false results- preform with rectal examination and biopsy
▪ May be elevated after ejaculation.
▪ Prostatic cancer
▪ Metastasis from prostatic cancer.
➢ fPSA/PSA:
▪ >25% not prostate cancer.
▪
Genitourinary tract tumor markers, clinical significance. Interpretation of results.
TESTICULAR GERM CELL (2)
Seminomatuos/ non-seminomatuos:
- non-seminomatuos grow faster
- early onset of cancer
- lower survival rate
➢ ßHCG:
▪ Increase in seminoma with no increase in AFP.
▪ Better TM then hCG.
▪ Never found in normal men.
▪ Testicular cancer.
▪ The amount correlate with tumor mass.
▪ Used in- diagnosis, staging, management and detection of reoccurrence
➢ AFP:
▪ Elevated in non- seminoma cancer with or without ßHCG
Genitourinary tract tumor markers, clinical significance. Interpretation of results.
URINARY BLADDER (3)
➢ Nuclear matrix protein (NMP22):
▪ + in 70% of bladder carcinoma.
▪ Not used for screening.
▪ Used for follow ups.
➢ Bladder tumor antigen (BTA):
▪ Elevated in urine in bladder tumors.
▪ High in kidney stones and UTI.
➢ Tissue polypeptide antigen (TPA):
▪ Marker of proliferation and to specific for tumors.
▪ Used for follow up.
▪ Bladder, breast, bronchial, cervical, ovarian and colorectal carcinoma.
Gynecological tumor markers: (3)
Ovarian carcinoma: (9) -Ca 125, -Ca 72-4, -LASA-P, -CEA, -TPA, -BRCA1, -BRCA2, -HE4, -SMRP Ovarian germ cells: (2) -hCG -AFP Gestational trophoblastic disease: (1) -hCG
Gynecological tumor markers: clinical significance. Interpretation of results.
OVARIAN (5)
➢ Ca 125:
▪ Not used as screening test.
▪ Endometrial, uterine tube and lung malignancies.
▪ Most used serum biomarker.
▪ Used for diagnosis, response to treatment and reoccurrence.
▪ Have high rate of false positive
➢ Human epididymis protein 4 (HE4):
▪ New biomarker for early diagnosis.
▪ High sensitivity and specificity.
➢ Mesothelin (SMRP):
▪ In combination with HE4 is god early diagnosis.
▪ Complimentary diagnostic tool with Ca 125.
▪ Detected in other body fluids not just blood.
▪ Ovarian cancer, pancreatic adenocarcinoma, mesotelioma.
➢ Ca 72-4:
▪ Detects stomach carcinoma
▪ If combined with Ca125- detects ovarian carcinoma.
➢ BRCA-1/2:
▪ Genetic marker.
▪ Human suppressor genes.
Gynecological tumor markers: clinical significance. Interpretation of results.
OVARIAN GERM CELLS (2)
Can be cancerous or non.
Birth defect.
➢ hCG:
▪produced in the placenta
▪ Germ cells tumors
▪ Choriocarcinomas
▪ Gestational trophoblastic disease.
▪ Correlates with staging and tumor mass.
▪ Have little clinical application in- breast, lung, prostate, kidney, pancreatic, bladder and gastrointestinal tract cancers
➢ BhCG:
▪ Better then hCG.
▪ Used in diagnosis and monitoring.
▪ False positive in vegetarians
▪ False negative due to drugs and fluids.
▪ Increased in- duodenal ulcer, cirrhosis, pregnancy.
Gynecological tumor markers: clinical significance. Interpretation of results.
GESTATIONAL THROMBOPLASTIC D. (1)
➢ hCG:
▪produced in the placenta
▪ Germ cells tumors
▪ Choriocarcinomas
▪ Gestational trophoblastic disease.
▪ Correlates with staging and tumor mass.
▪ Have little clinical application in- breast, lung, prostate, kidney, pancreatic, bladder and gastrointestinal tract cancers
Breast tumor markers, clinical significance. Interpretation of results.
(11)
- Ca 27.29,
- Ca 15-3,
- CEA,
- ER/PR,
- HER-2neu,
- CTC,
- BRCA1,
- BRCA2,
- PLP,
- uPA,
- PAI-1.
Breast tumor markers, clinical significance. Interpretation of results.
(7)
Ca15-3, Ca27.29:
▪ Not organ specific.
▪ Not sensitive and not specific.
▪ Used for- reoccurrence, monitoring and metastasis.
▪ High in metastatic breast caner and stage III,IV
➢ Choriembryionic antigen (CEA):
▪ Not specific for breast cancer.
▪ If combined with Ca15-3 à increase its sensitivity.
➢ Estrogen/progesterone receptors (ER/PR):
▪ If (+) cancer grow slowly.
▪ ER cancer is less aggressive.
▪ ER (+) à decreased risk for reoccurrence.
▪ Treatment is with hormones.
➢ Human epidermal growth factor receptor- 2 (HER-2):
▪ Oncogene.
▪ Used for diagnosis, treatment and prognosis.
▪ Give info only about primary breast cancer.
➢ BRCA-1/2:
▪ Increase risk of breast cancer in 90%.
▪ Men high risk for breast and prostate cnacer.
➢ Circulating tumor cells (CTC):
▪ Not specific.
▪ Found in peripheral blood, bone marrow and metastasis.
▪ Decreased survival.
➢ P53: tumor suppressor gene.
▪ Occur in 50% of patients.
▪ More aggressive.
Laboratory diagnostic of acute leukemias:
- Examination of the blood
- Decreased red cells, reticulocytes and platelets count
- WBC – varying from leukopenia to very high numbers
- Circulating blasts
Laboratory diagnostic of acute leukemia:
- Bone marrow aspirate microscopy
- Hypercellular
- Sheets of blasts replacing the normally maturing cells in the erythroid, myeloid, and megakaryocytic lines.
- Blasts >20%
Laboratory diagnostic of acute leukemias:
- Cytochemical stains (4)
Identification AML/ALL + sub-classification
- Peripheral blood films
- Bone marrow aspirates
- Lymph node / other tissue biopsies
- MPO- Myeloperoxidase activity.
Granules of neutrophils and eosinophils contain
myeloperoxidase.
Monocytic lysosomal granules are faintly positive.
Lymphocytes and nucleated red blood cells lack the enzyme. - SBB- Sudan black B stained lipids in black.
Myeloid line cells contains lipids.
Lymphoid line cells don’t have lipids. - PAS- Periodic acid-Schiff.
Stain dyes red glycogen and polysaccharides granules in blasts.
Myeloid blasts small granules.
Lymphoid blasts large granules. - ANAE- alphanaphthyl acetate esterase
Only monocytic line cells that stained brown.
Laboratory diagnostic of acute leukemias by:
FLOW CYTROMETRY
- Principles of method
Analyses physical and chemical characteristics of particles.
Cell components are fluorescently labeled and then excited by the laser to emit light at varying wavelengths.
- The light scatter measurements reflect the physical properties of the cells (cell size or internal complexities).
- The fluorescence data give information on the membrane or intracellular molecules (proteins, DNA) depending on the antibodies and dyes used for labeling the cells.
Laboratory diagnostic of acute leukemias by:
FLOW CYTROMETRY
- Samples
Fresh specimens for FC processing and analysis, 2 categories:
Liquid samples (3)
- peripheral blood
- bone marrow
- body fluids
Solid tissue (5)
- lymph nodes
- tonsils/ adenoids
- spleen
- bone marrow biopsies
- extranodal infiltrates
Laboratory diagnostic of acute leukemias by:
FLOW CYTROMETRY
- Possibility of diagnostic
Only for AML, ALL and CLL diagnostic.
Laboratory diagnostic of chronic leukemias:
- Examination of the blood
CBC and blood film show
⬆️ WBC (generally >25 x 10^9/L, often 100–300 x10^9/L)
(Mostly neutrophils and myelocytes; basophilia; sometimes eosinophilia)
➡️ Anaemia common
➡️ Platelets: N / ⬆️
⬇️NAP and ESR (in absence of secondary infection)
⬆️LDH and Urate
*Cytogenetic examination of blood or marrow for confirmatory t(9;22)
Laboratory diagnostic of chronic leukemias:
- Bone marrow aspirate microscopy
- Hypercellularity due to myeloid hyperplasia
(blasts 10% in accelerated phase; >20% blasts + promyelocytes = blast crisis) - TREPHINE useful to assess marrow fibrosis
- Cytogenetic examination of blood or marrow for confirmatory t(9;22)
Tumor markers: definition
Molecules, occurring in blood or tissue that are associated with cancer and whose measurement or identification is useful in patient diagnosis or clinical management.
Usually present at normal or very low levels in the blood of healthy persons.
If the substance in question is produced by the tumor, its levels will be increased.
